Towards the Production of mcl-PHA with Enriched Dominant Monomer Content: Process Development for the Sugarcane Biorefinery Context

The production of short-chain-length polyhydroxyalkanoates (scl-PHAs) in a sugarcane biorefinery setting has been demonstrated to be an effective strategy to reduce production costs. Medium-chain-length PHA (mcl-PHA) have elastomeric properties and are more suitable for high value-added applications, but its industrial production is not yet established. Mcl-PHA synthesis occurs via different metabolic routes and thus requires distinct microorganisms and substrates compared to scl-PHA. In the present study, sucrose-derived carbohydrates were evaluated as co-substrates for the production of mcl-PHA from decanoic acid (DA). Fermentation strategies were investigated to produce mcl-PHA with enriched dominant monomer content, which is desirable for commercial applications. The mcl-PHA production was investigated in carbon-limited, fed-batch fermentations with wild-type and β-oxidation knockout mutant strains of Pseudomonas putida KT2440. The experimental results demonstrated that a mixture of glucose and fructose was a suitable co-feed with DA for mcl-PHA production, yielding equivalent results to those obtained with starch-derived glucose, a more traditional feedstock for PHA production. The use of a β-oxidation-impaired strain was essential to attain high dominant monomer content. A near-homopolymeric mcl-PHA was produced under exponential feeding, containing 99 mol% of 3-hydroxydecanoate. This work demonstrates the potential for near-homopolymeric mcl-PHA production in a sugarcane biorefinery, using hydrolyzed sucrose and DA.